Fig. 4

TLR2 plays a key role in inducing G-CSF in GR-1-treated BMDM in a phagocytosis dependent manner. a Immortalized BMDM were treated with the actin polymerase inhibitor cytochalasin D (CytD) for 1 h and then exposed to live GR-1 (20 CFU/cell) or lipoteichoic acid (LTA; 10 μg/ml). b Primary BMDMs prepared from wild-type (WT), TLR2^−/−, NOD1^−/− and NOD2^−/− mice were treated with live GR-1 (20 CFU/cell), LTA (10 μg/ml) or Pam₃CSK₄ (PAM3; 1 μg/mL). a-b Production of TNF-α (in 4 h) and G-CSF (in 24 h) was measured from spent cell culture media using ELISA. Data shown as mean ± SEM [n ≥ 3]. c Primary BMDMs from wild-type (WT), TLR2^−/−, NOD1^−/− and NOD2^−/− mice were treated with live GR-1 (20 CFU/cell) for 15, 30 and 60 min. Phosphorylation of MAPKs (ERKs, p38 and JNKs), Akt and I-κB was analyzed by Western blots, using phospho- or β-actin antibodies. Bar graphs demonstrate band intensity quantification (ratio of protein phosphorylation to β-actin) using the NIH ImageJ program. Data shown as mean ± SEM [n ≥ 3]